Facile preparation of fibrin coated open tubular column for characterization of monoclonal antibody variants by capillary electrochromatography

2017 ◽  
Vol 140 ◽  
pp. 377-383 ◽  
Author(s):  
Xue Xiao ◽  
Wentao Wang ◽  
Yamin Zhang ◽  
Li Jia
Keyword(s):  
Monoclonal Antibody ◽  
Capillary Electrochromatography ◽  
Open Tubular Column ◽  
Facile Preparation

Functional Characterization of PM6/13, a β3-specific (GPIIIa/CD61) Monoclonal Antibody that Shows Preferential Inhibition of Fibrinogen Binding over Fibronectin Binding to Activated Human Platelets

1998 ◽  
Vol 79 (01) ◽  
pp. 177-185 ◽  
Author(s):  
Ashia Siddiqua ◽  
Michael Wilkinson ◽  
Vijay Kakkar ◽  
Yatin Patel ◽  
Salman Rahman ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Platelet Aggregation ◽  
Platelet Rich Plasma ◽  
Functional Characterization ◽  
Human Platelets ◽  
Western Blots ◽  
Activated Platelets ◽  
Reducing Conditions ◽  
Fibronectin Binding

SummaryWe report the characterization of a monoclonal antibody (MAb) PM6/13 which recognises glycoprotein IIIa (GPIIIa) on platelet membranes and in functional studies inhibits platelet aggregation induced by all agonists examined. In platelet-rich plasma, inhibition of aggregation induced by ADP or low concentrations of collagen was accompanied by inhibition of 5-hydroxytryptamine secretion. EC50 values were 10 and 9 [H9262]g/ml antibody against ADP and collagen induced responses respectively. In washed platelets treated with the cyclooxygenase inhibitor, indomethacin, PM6/13 inhibited platelet aggregation induced by thrombin (0.2 U/ml), collagen (10 [H9262]g/ml) and U46619 (3 [H9262]M) with EC50 = 4, 8 and 4 [H9262]g/ml respectively, without affecting [14C]5-hydroxytryptamine secretion or [3H]arachidonate release in appropriately labelled cells. Studies in Fura 2-labelled platelets revealed that elevation of intracellular calcium by ADP, thrombin or U46619 was unaffected by PM6/13 suggesting that the epitope recognised by the antibody did not influence Ca2+ regulation. In agreement with the results from the platelet aggregation studies, PM6/13 was found to potently inhibit binding of 125I-fibrinogen to ADP activated platelets. Binding of this ligand was also inhibited by two other MAbs tested, namely SZ-21 (also to GPIIIa) and PM6/248 (to the GPIIb-IIIa complex). However when tested against binding of 125I-fibronectin to thrombin stimulated platelets, PM6/13 was ineffective in contrast with SZ-21 and PM6/248, that were both potent inhibitors. This suggested that the epitopes recognised by PM6/13 and SZ-21 on GPIIIa were distinct. Studies employing proteolytic dissection of 125I-labelled GPIIIa by trypsin followed by immunoprecipitation with PM6/13 and analysis by SDS-PAGE, revealed the presence of four fragments at 70, 55, 30 and 28 kDa. PM6/13 did not recognize any protein bands on Western blots performed under reducing conditions. However Western blotting analysis with PM6/13 under non-reducing conditions revealed strong detection of the parent GP IIIa molecule, of trypsin treated samples revealed recognition of an 80 kDa fragment at 1 min, faint recognition of a 60 kDa fragment at 60 min and no recognition of any product at 18 h treatment. Under similar conditions, SZ-21 recognized fragments at 80, 75 and 55 kDa with the 55kDa species persisting even after 18 h trypsin treatment. These studies confirm the epitopes recognised by PM6/13 and SZ-21 to be distinct and that PM6/13 represents a useful tool to differentiate the characteristics of fibrinogen and fibronectin binding to the GPIIb-IIIa complex on activated platelets.

Immunochemical characterization of anti-islet cell surface monoclonal antibody from nonobese diabetic mice

Diabetes ◽  
1986 ◽  
Vol 35 (5) ◽  
pp. 517-522 ◽  
Author(s):  
J. Hari ◽  
K. Yokono ◽  
K. Yonezawa ◽  
K. Amano ◽  
S. Yaso ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Surface ◽  
Islet Cell ◽  
Diabetic Mice ◽  
Immunochemical Characterization ◽  
Nonobese Diabetic ◽  
Nonobese Diabetic Mice

scholarly journals Characterization of Native Reversible Self-Association of a Monoclonal Antibody Mediated by Fab-Fab Interaction

2020 ◽  
Vol 109 (1) ◽  
pp. 443-451 ◽  
Author(s):  
Lorenzo Gentiluomo ◽  
Dierk Roessner ◽  
Werner Streicher ◽  
Sujata Mahapatra ◽  
Pernille Harris ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Self Association

scholarly journals Characterization of ASP8374, a fully-human, antagonistic anti-TIGIT monoclonal antibody

2021 ◽  
pp. 100433
Author(s):  
Kenna Shirasuna ◽  
Gerald Koelsch ◽  
Cynthia Seidel-Dugan ◽  
Andres Salmeron ◽  
Philipp Steiner ◽  
...  
Keyword(s):  
Monoclonal Antibody

Characterization of a neutralizing anti-human galectin-1 monoclonal antibody with angioregulatory and immunomodulatory activities

Angiogenesis ◽  
2020 ◽  
Author(s):  
Juan M. Pérez Sáez ◽  
Pablo F. Hockl ◽  
Alejando J. Cagnoni ◽  
Santiago P. Méndez Huergo ◽  
Pablo A. García ◽  
...  
Keyword(s):  
Monoclonal Antibody
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